f10: Lung-derived gMDSCs induce metastatic growth of disseminated tumour cells.(a,b) We developed a mouse model that shows no metastatic growth when primary tumours were resected at 1 week post implantation (c) despite the existence of disseminated tumour cells in regional lymph nodes and lungs. (d,e) All mice develop metastasis when primary tumours were resected at 2 weeks post implantation. (f) Illustration of the experimental design. (g) Primary 4T1-Luci tumours were resected after 1 week post implantation and mice were followed up for metastatic growth by bioluminescence imaging (BLI). There was no metastatic growth up to week 11. (h) After resection of primary tumours, mice were injected (via tail vein) with tumour-derived mMDSCs as indicated and followed up by BLI without any metastatic growth. (i,j) Mice injected with lung-derived gMDSCs showed metastatic growth in three out of four mice. (k) Our findings suggest a spatiotemporal regulation of tumour plasticity by MDSC subsets in primary site and in distant organs as illustrated. Results are presented as mean±s.d. (five mice in each group). Scale bar, 50 μm.

Mentions:
To determine whether gMDSCs support the growth of already disseminated tumour cells, we developed a mouse model where orthotopically (fat pads) implanted tumours were resected at 1 week post implantation. Despite the presence of disseminated tumour cells in regional lymph nodes and lungs, there was no metastatic growth up to 12 weeks of follow up (Fig. 10a–c). In contrast, majority of animals developed metastasis when the primary tumours were resected at 2 weeks post implantation (Fig. 10d,e). First of all, these findings provided further evidence that infiltration of gMDSCs in the secondary organs is required for successful metastasis. As shown in Fig. 2, expansion and infiltration of gMDSCs in lungs occur at 2 weeks post implantation. Second, this model may offer a great utility for investigation of the disseminated tumour cells in the absence of primary tumours as shown in the experimental outline (Fig. 10f). We utilized this model to evaluate the functional role of gMDSCs. In three groups of mice, luciferase-tagged primary tumours were resected at 1 week post implantation. First control group were not treated thereafter resection (Fig. 10g), second group were injected twice with tumour-derived mMDSCs (250 K per mice by tail vein; Fig. 8h) and third group were injected twice with lung-derived gMDSCs (250 K per mice by tail vein; Fig. 10i) isolated from 4T1 tumour-bearing animals. First and second group of mice were followed up for metastatic growth by bioluminescence imaging (BLI) up to 11 weeks without any detectable metastasis (Fig. 10g,h). In contrast, 3 out of 4 mice injected with lung-derived gMDSCs developed metastasis (Fig. 8i,j). Collectively, our data suggest that dissemination and metastatic colonization/growth are two independent steps in the metastatic cascade and may be regulated by different subsets of MDSCs as depicted by the illustration of our working hypothesis (Fig. 10k).

f10: Lung-derived gMDSCs induce metastatic growth of disseminated tumour cells.(a,b) We developed a mouse model that shows no metastatic growth when primary tumours were resected at 1 week post implantation (c) despite the existence of disseminated tumour cells in regional lymph nodes and lungs. (d,e) All mice develop metastasis when primary tumours were resected at 2 weeks post implantation. (f) Illustration of the experimental design. (g) Primary 4T1-Luci tumours were resected after 1 week post implantation and mice were followed up for metastatic growth by bioluminescence imaging (BLI). There was no metastatic growth up to week 11. (h) After resection of primary tumours, mice were injected (via tail vein) with tumour-derived mMDSCs as indicated and followed up by BLI without any metastatic growth. (i,j) Mice injected with lung-derived gMDSCs showed metastatic growth in three out of four mice. (k) Our findings suggest a spatiotemporal regulation of tumour plasticity by MDSC subsets in primary site and in distant organs as illustrated. Results are presented as mean±s.d. (five mice in each group). Scale bar, 50 μm.

Mentions:
To determine whether gMDSCs support the growth of already disseminated tumour cells, we developed a mouse model where orthotopically (fat pads) implanted tumours were resected at 1 week post implantation. Despite the presence of disseminated tumour cells in regional lymph nodes and lungs, there was no metastatic growth up to 12 weeks of follow up (Fig. 10a–c). In contrast, majority of animals developed metastasis when the primary tumours were resected at 2 weeks post implantation (Fig. 10d,e). First of all, these findings provided further evidence that infiltration of gMDSCs in the secondary organs is required for successful metastasis. As shown in Fig. 2, expansion and infiltration of gMDSCs in lungs occur at 2 weeks post implantation. Second, this model may offer a great utility for investigation of the disseminated tumour cells in the absence of primary tumours as shown in the experimental outline (Fig. 10f). We utilized this model to evaluate the functional role of gMDSCs. In three groups of mice, luciferase-tagged primary tumours were resected at 1 week post implantation. First control group were not treated thereafter resection (Fig. 10g), second group were injected twice with tumour-derived mMDSCs (250 K per mice by tail vein; Fig. 8h) and third group were injected twice with lung-derived gMDSCs (250 K per mice by tail vein; Fig. 10i) isolated from 4T1 tumour-bearing animals. First and second group of mice were followed up for metastatic growth by bioluminescence imaging (BLI) up to 11 weeks without any detectable metastasis (Fig. 10g,h). In contrast, 3 out of 4 mice injected with lung-derived gMDSCs developed metastasis (Fig. 8i,j). Collectively, our data suggest that dissemination and metastatic colonization/growth are two independent steps in the metastatic cascade and may be regulated by different subsets of MDSCs as depicted by the illustration of our working hypothesis (Fig. 10k).